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TiO2 nanowire array substrate and preparation method and application thereof

A nanowire array and substrate technology, applied in biochemical equipment and methods, cell dissociation methods, tumor/cancer cells, etc., can solve the problems of impaired cell activity, time-consuming, expensive equipment, etc., and achieve high efficiency and specificity. The effect of capturing, increasing surface area, and improving capture efficiency

Pending Publication Date: 2021-08-24
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The third is through the TIO 2 Modification of degradable MnO on nanowire arrays 2 Particles for cell capture, and then use oxalic acid for MnO 2 Particle degradation is used to achieve the release of cells. This method can only achieve the overall release of cells, and the introduction of oxalic acid will lead to impaired cell activity.
The disadvantage of this method is that the preparation process of the gelatin substrate with cell topography is complicated and takes a lot of time
And because gelatin is easy to change state with the change of temperature, the prepared morphology is not easy to preserve
In addition, this method uses high-precision near-infrared light, and the equipment is expensive, which is not conducive to popularization

Method used

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  • TiO2 nanowire array substrate and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Preparation of antibody-modified TiO 2 Nanowire Array Substrate.

[0055] 1. Preparation of TiO 2 array of nanowires.

[0056] (1) Synthesis of TiO by hydrothermal method on the substrate 2 The nanowire array, the specific steps are as follows: (1) ultrasonically clean the FTO glass with acetone, absolute ethanol, and triple-distilled water for 15 minutes in sequence;

[0057] (2) Place the cleaned FTO glass in an oven at 65°C to dry;

[0058] (3) 30ml of concentrated hydrochloric acid with a concentration of 12mol / L, 30ml of deionized water and 1ml of tetrabutyl titanate were poured into the Erlenmeyer flask in sequence, and the reaction solution was obtained after mixing;

[0059] (4) Place the FTO glass dried in step (2) in the reactor, then pour the reaction solution obtained in step (3), then place the reactor at a temperature of 155° C., and react for 8 hours, namely have to.

[0060] 2. Apply gelatin.

[0061] (1) Doping gold nanorods in gelatin, the conce...

experiment example 2

[0070] Cell capture experiments.

[0071] 1. Set smooth glass group (SG), smooth glass group modified with antibody (SG / Anti) and TNA group as experimental controls;

[0072] 2, the TiO obtained in embodiment 1 2 The nanowire array substrate and the above three groups of experimental control substrates, a total of four substrates were co-incubated with MCF-7 cells;

[0073] 3. Specific incubation conditions: the concentration of MCF-7 cells is 10 5 cell / ml, incubate at room temperature for 1 hour.

[0074] The result is as figure 2 , image 3 shown, where figure 2 a, b, c, d in image 3 Smooth glass group (SG), TiO of unmodified antibody in 2 The nanowire array substrate (TNA), the smooth glass group (SG / Anti) modified with antibodies, and the substrate (TNA / Anti) obtained in Example 1 represent the cell density distribution under the fluorescence field of view. It can be seen from the statistical diagram that due to The joint effect of the morphology and the antibod...

Embodiment 3

[0076] Specific capture cell experiments.

[0077] 1, with the TiO that embodiment 1 obtains 2 Nanowire array substrates were co-incubated with MCF-7, A549, Hela, and WBC;

[0078] 2. The specific incubation conditions are: the concentrations of MCF-7, A549, Hela, and WBC cells are all 10 5 cell / ml, respectively and the TiO obtained in Example 1 2 The nanowire array substrate was incubated at room temperature for 1 hour.

[0079] The result is as Figure 4 As shown, it was found that the substrate modified with Anti-EpCAM antibody in Example 1 efficiently captured MCF-7 and A549 cells specifically expressing EpCAM antigen, but less captured Hela and WBC cells that did not express EpCAM antigen. This shows that the substrate has the ability to specifically recognize the captured cells.

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Abstract

The invention relates to a TiO2 nanowire array substrate and a preparation method and application thereof. The preparation method comprises the following steps: synthesizing a TiO2 nanowire array on a substrate by adopting a hydrothermal method; coating gelatin on the substrate on which the TiO2 nanowire array is synthesized; and modifying an antibody on the substrate coated with the gelatin. The TiO2 nanowire array substrate obtained by the preparation method can realize efficient specific capture of tumor cells, can selectively and locally release the captured cells, and enables the released cells to have high activity so as to facilitate subsequent biological analysis.

Description

technical field [0001] The invention belongs to the field of detection and release of circulating tumor cells, in particular to a TiO 2 Nanowire array substrate and its preparation method and application. Background technique [0002] Circulating tumor cells (CTCs) originate from primary or metastatic tumors, circulate in peripheral blood, and are the main form of tumor invasion and metastasis. Therefore, capturing and isolating circulating tumor cells from blood is crucial for early diagnosis and prognosis of cancer. However, the detection of CTCs still faces many difficulties and challenges. Since the concentration of CTCs is very low, typically, 1 mL of peripheral blood contains blood cells10 9 -10 10 1, but only about 1-100 CTCs are contained in 1 mL of peripheral blood, which requires the detection method used to efficiently and accurately detect very few target cells from a large number of cells. [0003] In response to this problem, there are four conventional so...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C03C17/42C03C17/25
CPCC12N5/0693C03C17/42C03C17/25C03C17/007C03C17/009C12N2509/00C03C2217/40C03C2217/70C03C2217/94C03C2218/111C03C2217/445C03C2217/479C03C2217/48
Inventor 李心磊范佳琪陈同生
Owner SOUTH CHINA NORMAL UNIVERSITY
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